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基于 rGONS 的生物传感器,可用于活细胞中 p53 和 p21 mRNA 的同时成像。

An rGONS-based biosensor for simultaneous imaging of p53 and p21 mRNA in living cells.

机构信息

College of Biology, Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, Hunan University, Changsha, 410082, China.

TCM and Ethnomedicine Innovation & Development International Laboratory, Innovative Material Medical Research Institute, School of Pharmacy, Hunan University of Chinese Medicine, Changsha, 410208, China.

出版信息

Talanta. 2019 Nov 1;204:20-28. doi: 10.1016/j.talanta.2019.05.087. Epub 2019 May 24.

Abstract

Hepatocarcinoma is the second leading cause of cancer-related death worldwide accompanied by the aberrant expression of many genes. Among of them, p53 and p21 genes played a vital role in the development of liver cancer. Thus, monitoring mRNA levels of the two markers is urgently needed for early diagnosis and understanding the molecular mechanism of hepatocarcinoma development. Herein, a functional nanosystem for in situ and real-time monitoring levels of p53 and p21 mRNA was constructed using reduced graphene oxide nanosheet assisted fluorescence probes. Comparing with traditional methods, the new method indicated high sensitivity and outstanding selectivity towards p53 and p21 mRNA assay in vitro. Moreover, the functional nanosystem was successfully used for monitoring dynamic change of mRNAs in HepG2 cells caused by cisplatin treatment, the reliability of which was confirmed by RT-PCR. In summary, this strategy provides a promising tool to reveal p53 and p21 mRNA regulatory process in cancer cells, which is practicable for drug screening and therapy evaluation on clinic.

摘要

肝癌是全球癌症相关死亡的第二大主要原因,伴随着许多基因的异常表达。其中,p53 和 p21 基因在肝癌的发展中起着至关重要的作用。因此,迫切需要监测这两种标志物的 mRNA 水平,以进行早期诊断和了解肝癌发展的分子机制。在这里,使用还原氧化石墨烯纳米片辅助荧光探针构建了用于原位和实时监测 p53 和 p21 mRNA 水平的功能纳米系统。与传统方法相比,该新方法在体外对 p53 和 p21 mRNA 测定表现出高灵敏度和出色的选择性。此外,该功能纳米系统成功地用于监测顺铂处理引起的 HepG2 细胞中 mRNA 的动态变化,其可靠性通过 RT-PCR 得到了证实。总之,该策略为揭示癌细胞中 p53 和 p21 mRNA 的调控过程提供了一种有前途的工具,在临床上可用于药物筛选和治疗评估。

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